A pathology review was conducted at the Department of Pathology, Faculty of Medicine, Comenius University, by a pathologist associated with the study. The study included tumor specimens corresponding to plasma samples from 78 patients. All specimens were classified according to the WHO Classification of 2004. The block containing the most representative part of the hematoxylin and eosin (H&E)-stained tumor was identified by microscopy and subsequently used for immunohistochemistry (IHC) analysis. Tissue microarray construction and immunohistochemical staining were performed as described previously (31 (link), 36 (link)). MMP1 was detected by primary rabbit polyclonal antibody against MMP1 (LSBio, MMP1, LS-B1229) diluted 1:40 in Dako REAL antibody diluent (Dako, Glostrup, Denmark), while MMP9 was detected by primary mouse monoclonal antibody against MMP9 (Abcam; MMP9 (SB15c); cat. no. ab51203) diluted 1:200 in Dako REAL antibody diluent (Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) as described previously (31 (link), 36 (link)).
The result of the immunohistochemical analyses was expressed by a weighted histoscore, evaluating both the percentage of positive cells and the staining intensity of the nuclei or cytoplasm as described previously (31 (link), 36 (link)).
The result of the immunohistochemical analyses was expressed by a weighted histoscore, evaluating both the percentage of positive cells and the staining intensity of the nuclei or cytoplasm as described previously (31 (link), 36 (link)).
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